A selective concentration-dependent dysrhythmogenic and antidysrhythmic action of prostaglandins E2, F2α and I2 (prostacyclin) on isolated rat hearts

Summary Prostaglandins (PGs) E₂, F₂, and I₂ were examined for their effects on the electrical and mechanical activities of isolated rat, rabbit and guinea-pig hearts. All PGs produced dysrhythmias in rat hearts at low concentrations only, while higher concentrations were antiarrhythmic. Guinea-pig hearts were less responsive while rabbit hearts were completely resistant.We thank the Manitoba Heart Foundation for financial support and Dr J. Pike (Upjohn, Kalamazoo, MI, USA) for gifts of prostaglandins. Dr M. Karmazyn is a Fellow of the Canadian Heart Foundation.     

Effects on prostaglandins F2α, I2, and indomethacin on isolated coronary arteries from healthy and alloxandiabetic dogs

Summary Contractile responses of isolated coronary arteries from healthy and alloxan-diabetic dogs to prostaglandin F₂ were enhanced by indomethacin and inhibited by prostaglandin I₂. The potentiation by indomethacin was more prominent in diabetic vessels than in normal arteries.     

Effects of PGE2 and PGF2α on the simulation by noradrenaline and oxytocin of human cervical muscle activity at term

Summary Cervical specimens were obtained by needle biopsy in connection with caesarean section at term pregnancy. The preparations were superfused in an organ chamber and contractions were registered isometrically. Prostaglandin (PG) E₂ and F₂ inhibited spontaneous contractions. The stimulatory action of noradrenaline was not influenced by PGF₂ but was reduced by PGE₂ whereas both PGs abolished the excitatory effect of oxytocin.     

Heavy incorporation of3H-prostaglandin F2α in the neoplastic cells as revealed by autoradiographic studies

Summary A marked uptake (9-fold) of the³H-PGF₂ was found specifically over heterochromatin in the nuclei of neoplastic cells. Lower but significant uptakes of³H-PGF₂ were also found in the nuclei of control epidermal cells, which indicate the presence of nuclear receptors in the epidermal neoplastic cells.     

An immunohistochemical and ultrastructural comparison of the effects of 2-bromo-α-ergocryptine on intrasellar and transplanted rat pituitaries

Summary Following 2 weeks of administration of 2-bromo--ergocryptine, a marked decrease was observed in prolactin immunoreactivity of the grafted pituitaries, whereas no reduction was noted in the intrasellar pituitaries. No evidence of crinophagy was revealed by electron microscopy in prolactin cells of 2-bromo--ergocryptine-treated rats.Acknowledgments. This work was supported in part by the Medical Research Council of Canada (grant MA-6349). The excellent technical assistance of Mrs Cynthia Edwards and secretarial help of Mrs Wanda Wlodarski are gratefully acknowledged.     

The effects of 9α-halohydrocortisones and of aldosterone on survival,growth and sodium-potassium excretion in adrenalectomized rats

Zusammenfassung Die Wirkungen von Aldosteron und 9-Halogenhydrocortison-azetaten auf das Überleben und Wachstum adrenalektomierter Ratten wurden studiert. Im Na/K-Ausscheidungstest zeigten sich 9-Chlor-hydrocortison-azetat bzw. 9-Fluor-hydrocortison-azetat 35-bzw. 10mal wirksamer als DCA und wiesen 1/3 bzw. 1/10 der Wirksamkeit von Aldosteron auf.     

Morphological and histochemical demonstration of a glycocalyx on the cell surface ofChattonella antiqua,a ‘naked flagellate’

Summary Electron microscopy demonstrated the presence of glycocalyx on the cell surface ofChattonella antiqua, which was previously believed to be naked. Histochemical analysis of this structure indicates that the glycocalyx consists of at least two different types of acidic complex carbohydrates, sulfated and nonsulfated, together with a neural carbohydrate-protein complex with 1,2-glycol groups and -D-mannosyl and -D-glucosyl residues.     

Folding and assembly defects of pyruvate dehydrogenase deficiency-related variants in the E1α subunit of the pyruvate dehydrogenase complex

The pyruvate dehydrogenase complex (PDC) bridges glycolysis and the citric acid cycle. In human, PDC deficiency leads to severe neurodevelopmental delay and progressive neurodegeneration. The majority of cases are caused by variants in the gene encoding the PDC subunit E1α. The molecular effects of the variants, however, remain poorly understood. Using yeast as a eukaryotic model system, we have studied the substitutions A189V, M230V, and R322C in yeast E1α (corresponding to the pathogenic variants A169V, M210V, and R302C in human E1α) and evaluated how substitutions of single amino acid residues within different functional E1α regions affect PDC structure and activity. The E1α A189V substitution located in the heterodimer interface showed a more compact conformation with significant underrepresentation of E1 in PDC and impaired overall PDC activity. The E1α M230V substitution located in the tetramer and heterodimer interface showed a relatively more open conformation and was particularly affected by low thiamin pyrophosphate concentrations. The E1α R322C substitution located in the phosphorylation loop of E1α resulted in PDC lacking E3 subunits and abolished overall functional activity. Furthermore, we show for the E1α variant A189V that variant E1α accumulates in the Hsp60 chaperonin, but can be released upon ATP supplementation. Our studies suggest that pathogenic E1α variants may be associated with structural changes of PDC and impaired folding of E1α.     

Effects of injury on the concentration ofα 1 andα 2 in the plasmas of male and female rats

Summary The effects of injury on the concentration of ₁-macroglobulin and ₂-macroglobulin in the plasmas of male and female rats has been investigated. At 5 days after injury to the male rats the ₁-macroglobulin concentration increased to 131% of its preinjury value. The ₂-macroglobulin concentration increased more rapidly to a maximum of 86 times its initial value. In the female rats ₂-macroglobulin increased only slightly and ₁-macroglobulin not at all.     

Efficient cell proliferation and predominant accumulation of ɛ-globin mRNA in human leukemic K562 cells which produce mostly Hb Gower 1

Summary Long-term cultures of K562(S) cells in 50–75 M hemin allow the selection of hemin-resistant K562 cells together with cells which proliferate efficiently while fully induced to express the human embryonic globin genes, as the hemoglobin Gower 1 (₂₂) is the predominant hemoglobin produced. Our experiments demonstrate that these K562 cells accumulate mostly -globin mRNA (-globin mRNA/-globin mRNA=2.9) suggesting that the control of hemoglobin expression is at a pretranslational level.We thank Dr Irene Bozzoni (Centro degli Acidi Nucleici, Università di Roma) for the pXCR7 probe. Address for reprint request: R.G. Centro Studi Biochimici sul Morbo di Cooley, Via Borsari 46, I-44100 Ferrara.     

15-Deoxy-Δ12,14-prostaglandin-J2 reveals a new pVHL-independent, lysosomal-dependent mechanism of HIF-1α degradation

Hypoxia-inducible factor-1α (HIF-1α) protein is degraded under normoxia by its association to von Hippel-Lindau protein (pVHL) and further proteasomal digestion. However, human renal cells HK-2 treated with 15-deoxy-Δ^12,14-prostaglandin-J₂ (15d-PGJ₂) accumulate HIF-1α in normoxic conditions. Thus, we aimed to investigate the mechanism involved in this accumulation. We found that 15d-PGJ₂ induced an over-accumulation of HIF-1α in RCC4 cells, which lack pVHL and in HK-2 cells treated with inhibitors of the pVHL-proteasome pathway. These results indicated that pVHL-proteasome-independent mechanisms are involved, and therefore we aimed to ascertain them. We have identified a new lysosomal-dependent mechanism of HIF-1α degradation as a target for 15d-PGJ₂ based on: (1) HIF-1α colocalized with the specific lysosomal marker Lamp-2a, (2) 15d-PGJ₂ inhibited the activity of cathepsin B, a lysosomal protease, and (3) inhibition of lysosomal activity did not result in over-accumulation of HIF-1α in 15d-PGJ₂-treated cells. Therefore, expression of HIF-1α is also modulated by lysosomal degradation.     

Effect of an ecdysteroid on DNA synthesis,DNA polymerase α and thymidine kinase activities ofDrosophila melanogaster cells in vitro

Summary 10^–7 M 20-OH-ecdysone treatment of a diploidDrosophila clone results in an inhibition of 60% of the DNA synthesis from 18 h of treatment on. After 48 h of hormone treatment the thymidine kinase activity is 70% inhibited; concomitantly a 60–70% lowering of the acid-soluble specific activity is observed. In the meantime the DNA polymerase activity is reduced.     

A human homolog of the yeast gene encoding tRNA 2′-phosphotransferase: cloning,characterization and complementation analysis

The Saccharomyces cerevisiae TPT1 gene plays a role in removing the 2-phosphate from ligated tRNA during the maturation of pre-tRNA. Here we reported the cloning and characterization of the human TRPT1 gene as a homolog of yeast TPT1. The TRPT1 gene is located at human chromosome 11q13 and encodes a polypeptide of 253 amino acids. BLAST searches with its amino acid sequence revealed the ubiquitous occurrence of TRPT1 homologs and their functional relationships with the presence of the DUF60/KptA domain. Northern analysis demonstrated that the gene is primarily expressed in heart and skeletal muscle, with lower or undetectable levels in other tissues studied. A plasmid-shuffling experiment showed that the human TRPT1 gene could complement the tpt1 mutation in S. cerevisiaeReceived 19 March 2003; received after revision 25 April 2003; accepted 22 May 2003     

A stepwise model of polyreactivity of the T cell antigen-receptor (TCR): its impact on the self–nonself discrimination and on related observations (receptor editing,anergy, dual receptor cells)

The existence of antigen-receptors, BCR, and T cell antigen-receptors, that are “polyreactive”, necessitates a rethinking of its effect on two problems faced by the “adaptive” immune system: the self (S)–nonself (NS) discrimination and the determination of effector class. Here, we will concentrate on the impact of polyreactivity on the S–NS discrimination. The anti-S cells interacting with S (i.e., responding to Signal 1) are on the pathway to inactivation. Before irreversibility sets in, these cells can be activated by a second signal (Signal 2) from an effector T-helper (eTh). As these polyreactive anti-S cells express anti-NS specificities, they can be activated by recognition of NS-epitopes in the host’s normal immunogenic load with the potential to result in autoimmunity. This problem is delineated using a discrete structural model, the corollaries of which are: (1) a two-step pathway for the purging of anti-S cells (i.e., the S–NS discrimination), and (2) defensible contexts within which to view the phenomena of receptor editing, anergy, and dual receptor cells.     

A simple synthesis of (E)-3-formylbut-2-enenitrile,and its use as a precursor of isotope-labelled zeatin and (±) dihydrozeatin

Summary (E)-3-Formylbut-2-enenitrile (4) is synthesized in 2 steps by reacting pyruvaldehyde dimethylacetal and acetonitrile in the presence of sodium methoxide, followed by acid hydrolysis to give 58% overall yield on distillation. The aldehyde4 can be stepwise and selectively reduced to (E)-3-hydroxymethylbut-2-enylamine (7a) in 37% total yield or exhaustivelyb reduced in 1 step to (±)-4-hydroxy-3-methylbutylamine (6) in 46% total yield. Compound7a and6 can be condensed with 6-chloropurine to give zeatin and (±)dihydrozeatin respectively. This provides a readily accessible method for isotope-labelled zeatin and its derivatives at side chain.Contribution No. 1064, Alberta Research CouncilProcess for the Preparation oftrans-3-formylbut-2-enenitrile S. Chen and J. MacTaggart, patent applicatioin registered in Canada and the United States of America.Acknowledgment. The author thanks Mr J. M. MacTaggart and Mrs C. M. Goulet for valuable experimental assistance and Dr R. M. Elofson for helpful discussions.     

Effects of ibotenic acid,quisqualic acid and their relatives on the excitability of an identifiable giant neurone of an African giant snail (Achatina fulica Férussac)

Summary An identifiable giant neurone, PON (periodically oscillating neurone), of Achatina fulica Férussac, inhibited by erythro--hydroxy-L-glutamic acid, was also inhibited by 2 relatives of -hydroxy glutamic acid, ibotenic acid and quisqualic acid. These substances similarly showed the effect on the neurone even in the chloride-free medium.The authors wish to thank Professor Emeritus T. Takemoto of Tohoku University, Dr H. Iwasaki of Takeda Chemical Co. and Professor C. H. Eugster of Zurich University for the donation of precious substances, and Miss H. Tamura for technical assistance.